Medulloblastoma subgroups remain stable across primary and metastatic compartments.
Medulloblastoma comprises four distinct molecular variants with distinct genetics, transcriptomes, and outcomes. Subgroup affiliation has been previously shown to remain stable at the time of recurrence, which likely reflects their distinct cells of origin. However, a therapeutically relevant question that remains unanswered is subgroup stability in the metastatic compartment. We assembled a cohort of 12-paired primary-metastatic tumors collected in the MAGIC consortium, and established their molecular subgroup affiliation by performing integrative gene expression and DNA methylation analysis. Frozen tissues were collected and profiled using Affymetrix gene expression arrays and Illumina methylation arrays. Class prediction and hierarchical clustering were performed using existing published datasets. Our molecular analysis, using consensus integrative genomic data, establishes the unequivocal maintenance of molecular subgroup affiliation in metastatic medulloblastoma. We further validated these findings by interrogating a non-overlapping cohort of 19 pairs of primary-metastatic tumors from the Burdenko Neurosurgical Institute using an orthogonal technique of immunohistochemical staining. This investigation represents the largest reported primary-metastatic paired cohort profiled to date and provides a unique opportunity to evaluate subgroup-specific molecular aberrations within the metastatic compartment. Our findings further support the hypothesis that medulloblastoma subgroups arise from distinct cells of origin, which are carried forward from ontogeny to oncology.


Illness perception in bulimia nervosa.
The study was designed to extend our understanding of illness perceptions in patients with bulimia nervosa (BN). Seventy-eight participants with BN or BN-type Eating Disorder Not Otherwise Specified (EDNOS-BN) completed the Revised Illness Perception Questionnaire (IPQ-R) (Moss-Morris et al., 2002). Clinical variables were also assessed. Participants experienced their ED as chronic, with serious consequences and high associated levels of anxiety and depression. The disorder was attributed primarily to psychological causes. The results indicate the perceived severity of BN, and high level of associated distress. These findings highlight the potential for targeting illness perceptions in treatment.


Relationship between radiologic morphology of the bone lengthening formation and its complications.
The objective was to study the different types of lengthened bone regeneration and their development during the various phases of the process to correlate them with patient factors and the surgical technique used, and to establish a possible relation between the development of the bone lengthening formation and the problems or complications. The authors studied the radiographs of a random group of 55 patients taken at three points during the course of treatment. The callus was classified with regard to its transverse diameter and the presence or absence of hypodense areas. The overall callus type was significantly influenced by the etiology, the osteotomy site, and the percentage lengthened. The percentage by which the limb was lengthened at the beginning of the process influences the overall morphology of the callus. Poor callus had been lengthened the most, atrophic callus the least. There was a correlation between the morphology of the overall callus at the end of treatment and the percentage lengthened, and between the percentage lengthened and the presence of bands at the end of treatment. The authors also found a significant correlation between age and the appearance of bands at the end of distraction. A central band was found among younger patients. The type of osteotomy affected the overall callus at the end of distraction and at the end of treatment and also influenced the transverse diameter. All the elongations with poor bone formation at the end of treatment were found to have undergone a diaphyseal osteotomy. The most common complication at the first follow-up and at the end of distraction was angulation. The diameter of the callus and the presence of bands at the end of treatment were significantly related to the complications. Fracture occurred in the first 2 weeks after removal of the external fixator in 88% of cases and in the third and fourth week in the rest. However, the segment had no significant influence on the appearance of complications. Lengthened callus with incomplete trabecular formations and hypodense areas at the end of the treatment has a high risk of fracture at the end of treatment. Callus with axial deviation, hypodense areas, or an insufficient transverse diameter during the lengthening procedure must be manipulated so that it reaches the maturing phase in better condition.


Blood-stained cerebrospinal fluid: traumatic puncture or haemorrhage?
Computed tomography fails to ascertain, or exclude, the presence of intracranial haemorrhage in a considerable number of cases, especially in subarachnoid haemorrhage and haemorrhagic infarcts. A number of other methods, including cerebrospinal fluid spectrophotometry and cytology have, therefore, been tested to define their diagnostic efficacy in 25 cases of confirmed intracranial haemorrhage and in 25 instances of blood-stained cerebrospinal fluid due to traumatic puncture. The combination of spectrophotometry and cytology proved to have a high diagnostic reliability. On the basis of these results a routine scheme of investigation is proposed.


Interleukin-18-induced human coronary artery smooth muscle cell migration is dependent on NF-kappaB- and AP-1-mediated matrix metalloproteinase-9 expression and is inhibited by atorvastatin.
The proliferation and migration of arterial smooth muscle cells (SMCs) are key events in the vascular restenosis that frequently follows angioplasty. Furthermore, SMC migration and neointimal hyperplasia are promoted by degradation of the extracellular matrix by matrix metalloproteinases (MMPs). Because we demonstrated previously that the proinflammatory and proatherogenic cytokine interleukin-18 (IL-18) stimulates SMC proliferation (Chandrasekar, B., Mummidi, S., Valente, A. J., Patel, D. N., Bailey, S. R., Freeman, G. L., Hatano, M., Tokuhisa, T., and Jensen, L. E. (2005) J. Biol. Chem. 280, 26263-26277), we investigated whether IL-18 induces SMC migration in an MMP-dependent manner and whether the 3-hydroxy-3-methylglutaryl-CoA reductase inhibitor atorvastatin can inhibit this response. IL-18 treatment increased both mRNA and protein expression of MMP9 in human coronary artery SMCs. Gel shift, enzyme-linked immunosorbent, and chromatin immunoprecipitation assays revealed a strong induction of IL-18-mediated AP-1 (c-Fos, c-Jun, and Fra-1) and NF-kappaB (p50 and p65) activation and stimulation of MMP9 promoter-dependent reporter gene activity in an AP-1- and NF-kappaB-dependent manner. Ectopic expression of p65, c-Fos, c-Jun, and Fra-1 induced MMP9 promoter activity. Specific antisense or small interfering RNA reagents for these transcription factors reduced IL-18-mediated MMP9 transcription. Furthermore, IL-18 stimulated SMC migration in an MMP9-dependent manner. Atorvastatin effectively suppressed IL-18-mediated AP-1 and NF-kappaB activation, MMP9 expression, and SMC migration. Together, our results indicate for the first time that the proatherogenic cytokine IL-18 induces human coronary artery SMC migration in an MMP9-dependent manner. Atorvastatin inhibits IL-18-mediated aortic SMC migration and has therapeutic potential for attenuating the progression of atherosclerosis and restenosis.


The Reticulocyte Hemoglobin Equivalent as a Screening Marker for Iron Deficiency and Iron Deficiency Anemia in Children.
Iron deficiency (ID) is one of the most common nutritional deficiencies in children worldwide and may result in iron deficiency anemia (IDA). The reticulocyte hemoglobin equivalent (Ret-He) provides information about the current availability of iron in erythropoiesis. This study aims to examine the validation of Ret-He as a screening marker for ID and IDA in children. Blood samples were retrospectively obtained from medical records. Anemia was defined according to the definition provided by the World Health Organization (WHO) for children. ID was defined by transferrin saturation (TSAT) < 20% and ferritin < 100 ng/mL. Children were classified into four groups: IDA, non-anemia iron deficiency (NAID), control and others. Out of 970 children, 332 (34.2%) had NAID and 278 (28.7%) presented with IDA. Analysis revealed that Ret-He significantly correlates with ferritin (rho = 0.41; p < 0.001), TSAT (rho = 0.66; p < 0.001) and soluble transferrin receptor (sTfR) (rho = -0.72; p < 0.001). For ROC analysis, the area under the curve (AUC) was 0.771 for Ret-He detecting ID and 0.845 for detecting IDA. The cut-off value for Ret-He to diagnose ID was 33.5 pg (sensitivity 90.7%; specificity 35.8%) and 31.6 pg (sensitivity 90.6%; specificity 50.4%) to diagnose IDA. The present study demonstrates Ret-He to be a screening marker for ID and IDA in children. Furthermore, Ret-He can be used as a single screening parameter for ID and IDA in children without considering other iron parameters. Economically, the use of Ret-He is highly relevant, as it can save one blood tube per patient and additional costs.


Perioperative management of colon cancer under Medicare risk programs.
To determine differences in perioperative care and outcomes for patients with colon cancer enrolled in Medicare health maintenance organizations compared with similar fee-for-service nonenrollees. Cross-sectional evaluation of hospital care and posthospital outcomes with data obtained from medical records. Nineteen health maintenance organizations representing all model types were selected from 12 states. The nonenrollee sample was drawn from the same areas. The sample included 412 enrollees and 401 nonenrollees, representing 65 hospitals for health maintenance organizations and 61 hospitals for fee-for-service. Nonenrollees were slightly older and had higher preoperative risk. Enrollees had shorter intervals between admission and surgery (enrollees, 1.55 days vs nonenrollees, 2.85 days). Differences in length of stay (enrollees, 10.9 days vs nonenrollees, 14.2 days) persisted even after controlling for preoperative health status. Differences in admissions to intensive care units (enrollees, 36.4% vs nonenrollees, 44.4%) were highly influenced by preoperative health status. Nonenrollees were more significantly likely to receive preoperative antibiotics, postoperative testing (eg, postoperative chest radiographs and electrocardiograms), and postoperative patient-controlled analgesia. Tumor staging was similar for both groups. Enrollees were more likely to be discharged home, while nonenrollees were more likely to be discharged to a nursing home. There were no significant differences in hospital deaths or postdischarge readmissions. Health maintenance organization enrollees with colon cancer received less clinical services of several types than similar patients in fee-for-service settings, had shorter hospital stays, and were less likely to be discharged to nursing homes. However, there was no evidence that they experienced different outcomes.


Characterization of feeding behavior traits in steers with divergent residual feed intake consuming a high-concentrate diet.
The objective of this study was to examine the differences in feeding behavior patterns of steers with divergent phenotypes for residual feed intake (RFI). Three trials were conducted with 508 Angus-based composite crossbred steers (body weight [BW] = 309 ± 57 kg) fed a high-concentrate diet in pens equipped with electronic feed bunks (GrowSafe System). Initial and final carcass ultrasound measurements (intra-muscular fat, backfat depth, and rib-eye area) were collected on days 0 and 70, and BW measured at 14-d intervals. Individual dry matter intake (DMI) and feeding behavior traits were collected for 70 d, and RFI calculated as the residual from the regression of DMI on average daily gain (ADG) and mid-test BW0.75. Steers were ranked by RFI and assigned to low-, medium-, and high-RFI classes based on ± 0.5 SD from the mean RFI within the trial. The feeding behavior traits evaluated in this study included frequency and duration of bunk visit (BV) and meal events, head-down (HD) duration, mean meal length, time-to-bunk interval, the maximum nonfeeding interval, and the day-to-day variation of these traits, defined as the root mean squared error (RMSE) from linear regression of each trait on the day of trial. Additionally, three ratio traits were evaluated: BV events per meal, HD duration per BV event, and HD duration per meal event. Low-RFI (feed-efficient) steers consumed 16% less (P < 0.01) DMI, while BW and ADG were not different compared with high-RFI steers. Low-RFI steers had 18% fewer and 21% shorter (P < 0.01) BV events, and 11% fewer and 13% shorter (P < 0.01) meal events per day compared with high-RFI steers. Furthermore, low-RFI steers exhibited less (P < 0.05) day-to-day variance in DMI, as well as in frequency and duration of BV and meal events and HD duration compared with high-RFI steers. Differences in feeding behavior traits due to RFI were minimally affected by covariate adjustment for DMI, indicating that steers with divergent RFI have distinct feeding behavior patterns that are largely independent of differences in DMI. These results suggest that feeding behavior traits may be useful biomarkers for the prediction of feed efficiency in beef cattle.


The effect of modes of delivery on infants' feeding practices.
Breast feeding has a great impact on the infant morbidity and mortality. According to Pakistan Demographic and Health survey (PDHS) infant mortality rate is 78 deaths per 1,000 live births. World Health Organization recommends that exclusive breast feeding for six months can decrease infant mortality rate by one-third. The objective of the study was to find out how the mode of delivery had impact on the practice of breast feeding. Data were collected for 2500 consecutive patients during a period of two years, and it was seen that maternal initiative to breast feed was low and problems with lactation were much more in cases delivering their babies via cesarean sections than those delivering theirs by normal delivery. Vaginal and cesarean section deliveries took place in 54% and 46% of the case, respectively. Thirty percent of the women studied felt that they had no problems regarding breastfeeding, but 70% of them had some sort of problems with breastfeeding their babies. When the women were matched for the mode of delivery, 58% of women who had breastfeeding problems belonged to the cesarean delivery group and 42% of complaining mothers were from women with normal delivery. The relative risk of having problems with breastfeeding for women subjected to cesarean was 1.38 and the odds ratio was 0.61. The findings of the present study indicate that more in depth counseling sessions are required for women undergoing operative delivery to improve breast feeding among them.


Spatial architecture of tumour-infiltrating lymphocytes as a prognostic parameter in resected non-small-cell lung cancer.
Tumour-infiltrating lymphocytes (TILs) are critically implicated in the clinical outcome and response to immunotherapy in non-small-cell lung cancer (NSCLC) patients. The functional competence of lymphocyte subpopulations is strongly conditioned by their spatial arrangement within the tumour immune microenvironment. The aim of this study was to determine whether the tissue localization of specific TIL subpopulations might have an impact on the risk of recurrence in surgically resected NSCLC. High-speed scanning of whole slide images was performed on immunohistochemically stained tissue sections from 97 NSCLC patients to assess the number and ratio of CD3+, CD8+ and PD-1+ T-lymphocytes. TIL distribution was computed considering the intratumoural (proximal or distal) and peripheral (invasive margin) localization as well as their location within the fibrotic tissue (immune excluded). The tumour proliferative index was assessed by Ki67 labelling. The impact of TILs number and distribution on clinical-pathological characteristics and outcomes were statistically analysed. High density and percentage of proximal CD8+ TILs and low PD-1-to-CD8 ratio had a positive impact on disease-free-survival (P = 0.03) and overall survival (P = 0.003). An inverse correlation was observed between the abundance of intratumoural CD8+ TILs carrying PD-1 inhibitory receptor and cancer cell proliferation. Cases with high compared to low fraction of immune excluded CD8+ TILs had significantly reduced 5-year overall survival (n events: 22 vs 12; P = 0.04) and disease-free survival (n events: 24 vs 16; P = 0.03) rates while the amount of CD3+ and CD8+ TILs located at the invasive margin had a favourable effect on the clinical course. Mapping TIL subpopulations may implement the definition of prognostic parameters in surgically resected NSCLC.


A coronary heart disease risk model for predicting the effect of potent antiretroviral therapy in HIV-1 infected men.
Many HIV-infected patients on highly active antiretroviral therapy (HAART) experience metabolic complications including dyslipidaemia and insulin resistance, which may increase their coronary heart disease (CHD) risk. We developed a prognostic model for CHD tailored to the changes in risk factors observed in patients starting HAART. Data from five cohort studies (British Regional Heart Study, Caerphilly and Speedwell Studies, Framingham Offspring Study, Whitehall II) on 13,100 men aged 40-70 and 114,443 years of follow up were used. CHD was defined as myocardial infarction or death from CHD. Model fit was assessed using the Akaike Information Criterion; generalizability across cohorts was examined using internal-external cross-validation. A parametric model based on the Gompertz distribution generalized best. Variables included in the model were systolic blood pressure, total cholesterol, high-density lipoprotein cholesterol, triglyceride, glucose, diabetes mellitus, body mass index and smoking status. Compared with patients not on HAART, the estimated CHD hazard ratio (HR) for patients on HAART was 1.46 (95% CI 1.15-1.86) for moderate and 2.48 (95% CI 1.76-3.51) for severe metabolic complications. The change in the risk of CHD in HIV-infected men starting HAART can be estimated based on typical changes in risk factors, assuming that HRs estimated using data from non-infected men are applicable to HIV-infected men. Based on this model the risk of CHD is likely to increase, but increases may often be modest, and could be offset by lifestyle changes.


Optimism as a mediator of the association between social support and peripartum depression among mothers of neonatal intensive care unit hospitalized preterm infants.
The birth of a preterm infant and the hospitalization in a neonatal intensive care unit (NICU) add extra functional and emotional demands to the typical transition to motherhood. Therefore, the prevalence of peripartum depression (PPD) among preterm mothers is especially high. As optimism was found to be a mediator of the association between social support and depression, the current study aimed to test this mediation in the population of NICU mothers. A sample of 128 Israeli mothers of 214 NICU hospitalized infants filled out self-report questionnaires regarding background variables, social support, optimism, and PPD symptoms. As hypothesized, optimism was found to partially mediate the association between social support and PPD symptoms among mother of preterm infants. The higher the perceived social support reported, the higher maternal optimism reported, and in turn, the lower PPD symptoms reported. In accordance with the Transactional Stress Theory, social support and optimism reflect maternal secondary appraisals that impact the way mothers of preterm infants cope with the challenging situation of a preterm birth and NICU hospitalization. Healthcare professionals should encourage preterm mothers' use of social support to increase optimism and reduce the risk of developing PPD.


The Effect of Intra-carpal Kirschner Wire Augmentation in Screw Fixation of Scaphoid - A Retrospective Cohort Study.
Scaphoid fractures are most often treated with a single headless compression screw. However, intercarpal Kirschner wire (K-wire) might be added to improve stability and fracture outcomes. This study will determine if there is a difference in treatment outcome (union rate and time to union) between scaphoid fracture fixations using a single headless compression screw with and without augmentation using a intracarpal intramedullary K-wire. We conducted a retrospective review of patients who underwent surgery for isolated scaphoid fractures over a 15 years period from December 2000 to December 2015. Only patients who underwent open surgery with bone grafting were included. They were divided into a group treated with a single screw fixation, and another group treated with screw and K-wire fixations. Forty-four (58.7%) patients had single screw fixation and 31 (41.3%) had screw augmented with K-wire fixation. The overall union rate was 88.0%, with an overall mean time to union of 5.3 months. There was no difference in union rate (p=0.84) and time to union (p=0.66) between the single screw group and combined screw and K-wire group. Univariate analysis found that older age (t=-2.11, p=0.04) had a significant effect on union rate. Regression model showed that age had a significant effect on months to union. In open fixation of scaphoid fractures with compression screw and bone grafting, union rate and time to union is comparable whether or not screw fixation was augmented with an intracarpal K-wire. There was no increased risk of complications associated with augmented screw. Age of patient affected time to union and union rate.


Prognostic value of metabolic tumor volume and total lesion glycolysis from ¹⁸F-FDG PET/CT in lymph node metastases and risk stratification of endometrial carcinoma.
To investigate the prognostic value of metabolic tumor volume (MTV) and total lesion glycolysis (TLG), measured by preoperative ¹⁸F-fluorodeoxyglucose positron emission tomography/computed tomography (¹⁸F-FDG PET/CT), in risk stratification of patients with endometrial carcinoma (EC). The patients with pathological diagnosis of EC who underwent preoperative ¹⁸F-FDG PET/CT imaging were retrospectively selected for analysis of the prognostic values of PET parameters in risk classification and lymph node metastases (LNMs). Receiver-operating-characteristic analysis was used to analyze the correlation of PET parameters cutoff values with deep myometrial invasion (MI), lymphovascular space involvement and LNM for prognostic values in risk stratification. The sensitivity, specificity, positive predictive value, negative predictive value and accuracy for detection of LNM are 83.3%, 99.7%, 90.9%, 99.5% and 99.2%, respectively. The MTV and TLG of primary lesion of EC in the patients with LNM are notably higher than those in patients without LNM, p<0.010. The MTV and TLG of the EC primary lesions in high-risk patients are significantly higher than those in low-risk patients (p<0.010), but the maximum standardized uptake value (SUVmax) is not. The MTV and TLG of primary lesions were superior to SUVmax for predicting of deep MI, LNM and high-risk of EC (p<0.005). MTV and TLG of primary lesions are more valuable in predicting risk stratification of EC patients. Preoperative ¹⁸F-FDG PET/CT imaging is useful in predicting the LNM of EC and may help guide pelvic lymphadenectomy to avoid unnecessary pelvic lymphadenectomy in EC patients with low-risk stratification.


Effect of a Perioperative Educational Video in Patients Undergoing Mohs Reconstruction: A Randomized Clinical Trial.
Skin cancer is the most common type of cancer in the United States, and most are treated with Mohs micrographic surgery (MMS) by fellowship-trained dermatologic surgeons. Complex reconstruction in cosmetically and functionally sensitive areas often requires a plastic surgery consult. The uncertainty regarding reconstructive options and cosmetic appearance is difficult emotionally and cognitively for patients. To improve patient understanding about MMS reconstruction with plastic surgeons, we investigated the utility of a multimedia presentation during perioperative education. A randomized clinical trial was conducted at a hospital-affiliated outpatient MMS clinic and surgery center in the United States. Patients in the intervention arm were given a tablet or Web site link to watch a multimedia presentation about reconstruction after MMS. All patients were given a questionnaire about their understanding of MMS reconstruction and satisfaction with the reconstructive care they received. A total of 46 patients were recruited for the study and completed the questionnaire. Of these patients, 22 were randomized to the test intervention arm, and the remaining 24 were in the control group. Among participants in the test intervention arm, feedback regarding the platform was positive with more than 90% of patients reporting that the video was engaging, useful, and easy to understand. Patient understanding and satisfaction scores in the test intervention arm ranked higher across all domains than those of the control group. This randomized clinical trial showed increases in patient understanding and satisfaction among patients undergoing MMS reconstruction with the implementation of a perioperative educational video.


Does Perceptual Simulation Explain Spatial Effects in Word Categorization?
In three experiments we investigated the origin of the effects of the compatibility between the typical location of entities denoted by written words (e.g., "up" for eagle and "down" for carpet) and either the actual position of the words on the screen (e.g., upper vs. lower part of the screen), or the response position (e.g., upper- vs. lower- key presses) in binary categorization tasks. Contrary to predictions of the perceptual simulation account (Barsalou, 1999), conceptual spatial compatibility effects observed in the present study (faster RTs when the typical position of the stimulus referent in the real word was compatible with either the stimulus or response physical position) seem to be independent of whether there was an overlap between simulated processes possibly triggered by the presented stimulus and sensory-motor processes actually required by the task. Rather, they appear to depend critically on whether the involved stimulus and/or response dimensions had binary, variable (vs. fixed) values. Notably, no stimulus-stimulus compatibility effect was observed in Experiment 3, when the stimulus physical position was presented in a blocked design (i.e., it was kept constant within each block of trials). In contrast, in all three experiments, a compatibility effect between response position and another (non-spatial) conceptual dimension of the stimulus (i.e., its semantic category) was observed (i.e., an effect analogous to the MARC [linguistic markedness of response codes] effect, which is usually observed in the number domain; Nuerk et al., 2004). This pattern of results is fully accounted for by the polarity principle, according to which these effects originate from the alignment of the polarities of either different stimulus dimensions or stimulus and response dimensions.


The suprapharmacologic dosing of antithrombin concentrate for Staphylococcus aureus-induced disseminated intravascular coagulation in guinea pigs: substantial reduction in mortality and morbidity.
An animal model of gram-positive septicemia was developed to evaluate the effects of antithrombin (AT) concentrates on morbidity, mortality, and laboratory consequences of disseminated intravascular coagulation (DIC). DIC was induced in guinea pigs by infusing Staphylococcus aureus (SA) isolated from blood cultures of patients with DIC (DIC-SA) or without DIC (non-DIC-SA). The non-DIC-SA animals and animals infused with sterile saline served as controls. Varying doses of AT were administered either 30 minutes or 24 hours after infusion of SA. DIC was confirmed within 4 hours by changes in prothrombin time, activated partial thromboplastin time, fibrinogen, fibrinogen-fibrin degradation products, and AT activity. Clinical bleeding was also evident. Mortality of untreated DIC-SA animals was 36% within 24 hours and up to 75% by 72 hours. Intervention with any dose of AT between 125 and 1,000 IU/kg 30 minutes after DIC-SA infusion was associated with 100% survival (P < or = .05 in the 250 IU/kg group) and sustained increases in AT activity and fibrinogen concentrations (P < or = .05). When AT was administered in combination with low molecular weight heparin (LMWH) or if LMWH was adminstered alone, mortality from DIC-SA was slightly, but not significantly reduced compared with untreated DIC-SA. Gross hemorrhage was observed premortem and at autopsy in all of the DIC-SA animals but in substantially fewer animals that received AT (P < or = .001 in the 250, 500, and 1,000 IU/kg groups). In contrast, groups treated with LMWH, alone or with AT, experienced hemorrhage and appeared to develop pathologic DIC. Fibrin formation in end-organs was detected in all guinea pigs in the untreated DIC-SA group and in the groups treated with 125 IU/kg AT and LMWH alone. AT doses between 250 and 1,000 IU/kg administered 30 minutes after DIC-SA infusion prevented fibrin formation in end-organs (P < or = .001 in the 250 and 1,000 IU/kg groups). AT administered 24 hours after DIC-SA could not reverse pre-existing histopathologic evidence of DIC but favorably affected survival, which reached statistical significance in the 1,000 IU/kg AT group (P < or = .025). In summary, suprapharmacologic doses of AT concentrate significantly decreased morbidity and mortality and ameliorated adverse changes in laboratory measures induced by DIC-SA in this guinea pig model and were not associated with untoward hemorrhagic complications. These findings provide justification for studying the use of AT therapy in patients with DIC-SA.


Polycystic Ovary Syndrome, Subfertility and Vitamin D Deficiency.
Polycystic ovarian syndrome (PCOS) is the most common endocrine female disorder, affecting 4-18% women of reproductive age. The prevalence of PCOS in South Asian women, especially in Pakistani women, is much higher (52%) as compared to white population (20 - 25% in UK). In Pakistan, vitamin D status displays 31.2% vitamin D insufficiency, 53.5% vitamin D deficiency, and 15.3% normal vitamin D values. Vitamin D deficiency, though very common in the general population, is even more prevalent in PCOS patients, seen in approximately 67-85%, which further correlates to their related comorbidities. A lot of researches have to be directed to examine the association among the PCOS and vitamin D, which may display monitoring role in several symptoms related to PCOS, such as ovulatory dysfunction, endocrine disruption, and insulin resistance. Vitamin D supplementation can, therefore, be employed to improve the metabolic and endocrine disorders; especially improving hormonal profile, oxidative stress, and ovulation outcome in PCOS patients. Key Words: Vitamin D, Infertility, Ovulatory Dysfunction, Insulin Resistance.


Phenethyl isothiocyanate inhibits angiogenesis in vitro and ex vivo.
Previous studies, including those from our laboratory, have revealed that phenethyl isothiocyanate (PEITC), a constituent of many edible cruciferous vegetables, not only affords significant protection against chemically induced cancer in animal models but also inhibits growth of cancer cells in culture and in vivo by causing cell cycle arrest and apoptosis induction. We now report a novel response to PEITC involving inhibition of angiogenesis in vitro and ex vivo at pharmacologically achievable concentrations. The PEITC treatment caused a decrease in survival of human umbilical vein endothelial cells (HUVEC) in a concentration- and time-dependent manner. The capillary-like tube structure formation (in vitro neovascularization) and migration (invasion potential) by HUVEC was also inhibited significantly in the presence of PEITC at pharmacologically relevant concentrations (<1 mumol/L). The PEITC-mediated inhibition of angiogenic features of HUVEC in vitro was associated with suppression of vascular endothelial growth factor (VEGF) secretion, down-regulation of VEGF receptor 2 protein levels, and inactivation of prosurvival serine-threonine kinase Akt. The PEITC treatment reduced migration by PC-3 human prostate cancer cells, which correlated with inactivation of Akt and suppression of VEGF, epidermal growth factor (EGF), and granulocyte colony-stimulating factor (G-CSF) secretion. The PEITC-mediated inhibition of PC-3 cell migration was statistically significantly attenuated by ectopic expression of constitutively active Akt. Most importantly, PEITC treatment inhibited ex vivo angiogenesis as revealed by chicken egg chorioallantoic membrane assay. In conclusion, the present study suggests that inhibition of angiogenesis may be an important mechanism in cancer chemoprevention by PEITC.


Challenges for nanomechanical sensors in biological detection.
Nanomechanical biosensing relies on changes in the movement and deformation of micro- and nanoscale objects when they interact with biomolecules and other biological targets. This field of research has provided ever-increasing records in the sensitivity of label-free detection but it has not yet been established as a practical alternative for biological detection. We analyze here the latest advancements in the field, along with the challenges remaining for nanomechanical biosensors to become a commonly used tool in biology and biochemistry laboratories.


Abnormal mitochondrial genomes in yeast restored to respiratory competence.
When crosses are performed between newly arisen, spontaneous petite mutants of Saccharomyces cerevisiae, respiratory competent (restored) colonies can form. Some of the restored colonies are highly sectored and produce large numbers of petite mutants. The high-frequency petite formation trait is inherited in a non-Mendelian manner, and elimination of mitochondrial DNA from these strains results in the loss of the trait. These results indicate that abnormal mitochondrial genomes are sometimes formed during restoration of respiratory competence. It is hypothesized that these abnormalities result either from recombination between mitochondrial DNA fragments to produce molecules having partial duplications contained on inverted or transposed sequences, or else recombinational "hot spots" have been expanded.


Effects of musical expertise on oscillatory brain activity in response to emotional sounds.
Emotions can be conveyed through a variety of channels in the auditory domain, be it via music, non-linguistic vocalizations, or speech prosody. Moreover, recent studies suggest that expertise in one sound category can impact the processing of emotional sounds in other sound categories as they found that musicians process more efficiently emotional musical and vocal sounds than non-musicians. However, the neural correlates of these modulations, especially their time course, are not very well understood. Consequently, we focused here on how the neural processing of emotional information varies as a function of sound category and expertise of participants. Electroencephalogram (EEG) of 20 non-musicians and 17 musicians was recorded while they listened to vocal (speech and vocalizations) and musical sounds. The amplitude of EEG-oscillatory activity in the theta, alpha, beta, and gamma band was quantified and Independent Component Analysis (ICA) was used to identify underlying components of brain activity in each band. Category differences were found in theta and alpha bands, due to larger responses to music and speech than to vocalizations, and in posterior beta, mainly due to differential processing of speech. In addition, we observed greater activation in frontal theta and alpha for musicians than for non-musicians, as well as an interaction between expertise and emotional content of sounds in frontal alpha. The results reflect musicians' expertise in recognition of emotion-conveying music, which seems to also generalize to emotional expressions conveyed by the human voice, in line with previous accounts of effects of expertise on musical and vocal sounds processing.


Influence of nutritional status on growth hormone-dependent circulating somatomedin-C activity in mature sheep.
The effect of daily administration of ovine GH for a period of 4 weeks on somatomedin-C biological activity in plasma was investigated in mature Merino sheep fed a maintenance energy intake (low plane; LP) or 1.6 times this amount (high plane; HP). The GH treatment resulted in a significant (P less than 0.05) increase in plasma GH levels in blood samples collected 23.5 h after each daily injection in both LP and HP groups. Plasma concentrations of somatomedin-C activity and insulin were significantly stimulated to a maximum level by the third GH injection and remained at this level for 7 days. Subsequently, circulating levels of both hormones fell to 40-50% of the peak response to GH and returned to basal levels within 48 h of the cessation of GH injections. In the HP group the response of plasma insulin and somatomedin-C activity to GH injection was greater than in the LP group.


Complex regional pain syndrome in children: incidence and clinical characteristics.
To study the clinical and epidemiological characteristics of complex regional pain syndrome (CRPS) in children. All children and adolescents under 16 years of age with a new diagnosis of CRPS who were reported to the Scottish Paediatric Surveillance Unit were included. Patients' recruitment ran between 1 November 2011 and 31 October 2015. Information was collected on patients' demography, clinical features, investigations, management and impact of disease on child and family. The diagnosis of CRPS was made on fulfilling the clinical criteria of the International Association for the Study of Pain. 26 cases of CRPS were reported over 4 years, giving a minimum estimated incidence of 1.16/100 000 (95% CI 0.87 to 1.44/100 000) children 5-15 years of age. Nineteen patients were female (73%) and mean age at diagnosis was 11.9 (range 5.5-15.4 years). The median interval between onset of symptoms and diagnosis was 2 months (range 1-12). The majority of children have single site involvement, with legs been more often affected than arms and the right side is more often affected than the left. There was a clear trauma at onset of the illness in 19 children and possible nerve injury in one. All investigations were normal and several treatment modalities were used with variable success. The disease had significant impacts on the patients' education and family lives. The estimated incidence of CRPS is 1.2/100 000 children 5-15 years old. The diagnosis of CRPS is often delayed. CRPS has a significant impact on children and their families.


Fundamental gene network rewiring at the second order within and across mammalian systems.
Genetic or epigenetic events can rewire molecular networks to induce extraordinary phenotypical divergences. Among the many network rewiring approaches, no model-free statistical methods can differentiate gene-gene pattern changes not attributed to marginal changes. This may obscure fundamental rewiring from superficial changes. Here we introduce a model-free Sharma-Song test to determine if patterns differ in the second order, meaning that the deviation of the joint distribution from the product of marginal distributions is unequal across conditions. We prove an asymptotic chi-squared null distribution for the test statistic. Simulation studies demonstrate its advantage over alternative methods in detecting second-order differential patterns. Applying the test on three independent mammalian developmental transcriptome datasets, we report a lower frequency of co-expression network rewiring between human and mouse for the same tissue group than the frequency of rewiring between tissue groups within the same species. We also find second-order differential patterns between microRNA promoters and genes contrasting cerebellum and liver development in mice. These patterns are enriched in the spliceosome pathway regulating tissue specificity. Complementary to previous mammalian comparative studies mostly driven by first-order effects, our findings contribute an understanding of system-wide second-order gene network rewiring within and across mammalian systems. Second-order differential patterns constitute evidence for fundamentally rewired biological circuitry due to evolution, environment or disease. The generic Sharma-Song test is available from the R package 'DiffXTables' at https://cran.r-project.org/package=DiffXTables. Other code and data are described in Section 2. Supplementary data are available at Bioinformatics online.


Surface Engineering Based on Conductive Agent Dispersion Uniformity: Strategies toward Performance Consistency of Lithium-Ion Batteries.
The consistency of lithium-ion battery performance is the key factor affecting the safety and cycle life of battery packs. Surface engineering of electrodes in production processes plays an important role in improving the consistency of battery performance. In this study, the drying process in the electrode manufacturing process is studied as the effect on surface engineering of the electrode materials, with consideration on impacting the battery performance. Specifically, the solid content of the slurry and drying temperature are considered to be the two factors that affect conductive agent dispersion uniformity in the porous electrodes. To achieve surface engineering on the dispersion uniformity of the conductive agent, the optimal processing parameters can be obtained by adjusting the temperature and solid content of the slurry. The mechanism of dispersion uniformity of the conductive agent is mainly related to the polyvinylidene fluoride grid structure. In the manufacturing of lithium-ion batteries, the electrode coated with 66% solid slurry and dried at 90-100 °C presents stable energy storage performance, which is beneficial to maintain the stable performance of the battery pack in the application.


A systematic review of predictors of maintenance of normotension after withdrawal of antihypertensive drugs.
The identification and treatment of hypertension in the general community has contributed to the reduction in strokes and coronary heart disease observed during the past 30 years. However, concerns have arisen that some patients may be receiving unnecessary antihypertensive drug therapy leading to wasted resources and the potential for adverse drug effects. Once therapy has been started, treating physicians have difficulty in selecting patients for withdrawal and have concerns regarding patient safety and their own legal liability. This study reviews and consolidates information from published studies to identify known predictors of the successful maintenance of normotension after antihypertensive drug withdrawal. The predictors were identified by determining the proportion of subjects with various baseline characteristics who remained normotensive while off medication for at least 12 months. From these data we have developed a clinical algorithm to help identify patients in whom antihypertensive drug withdrawal might be considered. This may assist primary care physicians in achieving successful withdrawal of antihypertensive therapy among selected hypertensive patients. The most consistent predictors identified were blood pressure (BP) (lower pretreatment, on treatment, and after withdrawal), nature of pharmacotherapy (fewer agents and lower dose), and preparedness to accept dietary intervention (weight and sodium reduction). On the basis of this information, a trial of withdrawal of antihypertensive medication might be recommended for patients who have mildly elevated, uncomplicated BP that is well controlled on a single agent, and who are motivated and likely to accept lifestyle changes.


Development of a German version of the Oswestry Disability Index. Part 2: sensitivity to change after spinal surgery.
When functional scales are to be used as treatment outcome measures, it is essential to know how responsive they are to clinical change. This information is essential not only for clinical decision-making, but also for the determination of sample size in clinical trials. The present study examined the responsiveness of a German version of the Oswestry Disability Index version 2.1 (ODI) after surgical treatment for low back pain. Before spine surgery 63 patients completed a questionnaire booklet containing the ODI, along with a 0-10 pain visual analogue scale (VAS), the Roland Morris disability questionnaire, and Likert scales for disability, medication intake and pain frequency. Six months after surgery, 57 (90%) patients completed the same questionnaire booklet and also answered Likert-scale questions on the global result of surgery, and on improvements in pain and disability. Both the effect size for the ODI change score 6 months after surgery (0.87) and the area under the receiver operating characteristics (ROC) curve for the relative improvement in ODI score in relation to global outcome 6 months after surgery (0.90) indicated that the ODI showed good responsiveness. The ROC method revealed that a minimum reduction of the baseline (pre-surgery) ODI score by 18% (equal to a mean 8-point reduction in this patient group) represented the cut-off for indicating a "good" individual outcome 6 months after surgery (sensitivity 91.4% and specificity 82.4%). The German version of the ODI is a sensitive instrument for detecting clinical change after spinal surgery. Individual improvements after surgery of at least an 18% reduction on baseline values are associated with a good outcome. This figure can be used as a reliable guide for the determination of sample size in future clinical trials of spinal surgery.


Umbilical cord blood derived endothelial progenitor cells for tissue engineering of vascular grafts.
A substantial limitation regarding present pediatric cardiac surgery is the lack of appropriate materials for the repair of congenital defects. To address this shortcoming, tissue engineering is a scientific field that aims at in vitro fabrication of living autologous grafts with the capacity of growth, repair, and regeneration. Here we focused on tissue engineered vascular grafts using human umbilical cord blood derived endothelial progenitor cells (EPCs), as a noninvasive cell source for pediatric applications. EPCs were isolated from 20 ml fresh human umbilical cord blood by Ficoll gradient centrifugation and cultured in endothelial basal medium containing growth factors. After proliferation and differentiation cells were analyzed by immunohistochemistry and seeded onto three-dimensional (3D) biodegradable vascular scaffolds (porosity > 95%, n = 22). Twenty-four hours after seeding the vascular grafts were positioned into a pulse-duplicator-in vitro system and grown for 48 hours under biomimetic conditions. A second group was grown 6 days statically and an additional 6 days biomimetically. Controls were cultured statically. Analysis of the grafts included immunohistochemistry, histology, and scanning electron microscopy. Preseeding differentiated EPCs indicated constant endothelial phenotypes including acetylated low-density lipoprotein, cluster of differentiation 31, von Willebrand factor, and endothelial nitric oxide synthetase. Seeded EPCs established favorable cell-to-polymer attachment and proliferation into the 3D tubular scaffolds. Both conditioned and static cellular constructs demonstrated positive staining for cluster of differentiation 31, von Willebrand factor, and expression of endothelial nitric oxide synthase. Human umbilical cord derived EPCs indicated exceptional growth characteristics used for tissue engineering of vascular grafts. These cells demonstrated a constant endothelial phenotype and related functional features. Based on these results EPCs seem to be a promising autologous cell source with regard to cardiovascular tissue engineering, particularly for the repair of congenital defects.


Caregiver burden among informal caregivers in the largest specialized palliative care unit in Malaysia: a cross sectional study.
Informal caregivers (IC) are often overshadowed by the attention required by the terminally ill. This study aims to reveal the estimated proportion of caregiver burden, psychological manifestations and factors associated with caregiver burden among IC in the largest specialized Palliative Care Unit (PCU) in Malaysia. This was a cross-sectional study involving IC attending a PCU. Caregiver burden and psychological manifestations were measured using previously translated and validated Zarit Burden Interview and DASS-21 questionnaires respectively. Two hundred forty-nine samples were selected for analysis. The mean ZBI score was 23.33 ± 13.7. About half of the population 118(47.4%) was found to experienced caregiver burden whereby majority have mild to moderate burden 90(36.1%). The most common psychological manifestation among IC is anxiety 74(29.7%) followed by depression 51(20.4%) and stress 46(18.5%). Multiple logistic regression demonstrated that women who are IC to patients with non-malignancy were less likely to experience caregiver burden. IC who were highly educated and spent more than 14 h per day caregiving were at least twice likely to experience caregiver burden. Finally, those with symptoms of depression and anxiety were three times more likely to suffer from caregiver burden. Caregiver burden among IC to palliative patients is prevalent in this population. IC who are men, educated, caregiving for patients with malignancy, long hours of caregiving and have symptoms of depression and anxiety are at risk of developing caregiver burden. Targeted screening should be implemented and IC well-being should be given more emphasis in local policies.


Intranasal oxytocin modulates EEG mu/alpha and beta rhythms during perception of biological motion.
Oxytocin (OT) plays a determining role in social and pair bonding in many vertebrates and increasing evidence suggests it is a social hormone also in humans. Indeed, intranasal administration of OT modulates several social cognitive processes in humans. Electrophysiological studies in humans associated the suppression of EEG in the mu/alpha and beta bands with perception of biological motion and social stimuli. It has been suggested that mu and beta suppression over sensory-motor regions reflects a resonance system in the human brain analogous to mirror neurons in the monkey. We therefore hypothesized that OT, a social hormone, would enhance this suppression, hence, for the first time, link the action of this neuropeptide with a human correlate of mirror neuron activity. Twenty-four students were administered 24 IU of OT or placebo intranasally in a robust, double-blind within-subject design. 45 min later participants were shown a point-light display of continuous biological motion of a human figure's walk. In the 8-10 Hz (low alpha/mu band) and in the 15-25 Hz beta band, a significant main effect of treatment showed that suppression was significantly enhanced in the OT versus the placebo conditions and that this suppression was widespread across the scalp. These results are a first step linking OT to the modulation of EEG rhythms in humans, suggesting that OT may have a role in allocating cortical resources to social tasks partly mediated by mirror neuron activity.


Comparison of the RNA-based EndoPredict multigene test between core biopsies and corresponding surgical breast cancer sections.
This study compared the perfomance of the RNA-based EndoPredict multigene test on core biopsies and surgical breast cancer specimens and analysed the influence of biopsy-induced tissue injuries on the test result. 80 formalin-fixed paraffin-embedded samples comprising paired biopsies and surgical specimens from 40 ER-positive, HER2-negative patients were evaluated. Total RNA was extracted and the EndoPredict score was determined. RNA yield was considerably lower in core biopsies, but sufficient to measure the assay in all samples. The EndoPredict score was highly correlated between paired samples (Pearson r=0.92), with an excellent concordance of classification into a low or high risk of metastasis (overall agreement 95%). The measurements are comparable between core biopsies and surgical sections, which suggest that the EndoPredict assay can be performed on core biopsy tissue. Inflammatory changes induced by presurgical biopsies had no significant effect on the RNA-based risk assessment in surgical specimens.


Suicide, assisted suicide, and medical illness.
Medical illness is an important part of the motivation for suicide--a significant factor in about 50% of suicides in patients over the age of 50 years and 70% of suicides in patients over the age of 70 years. Depression, anxiety, and ambivalence about dying characterize both medical patients who attempt suicide and those who request assisted suicide. When the physical and psychological sources of the desperation that underlies requests for assisted suicide are addressed, the desire for death diminishes and patients are usually grateful for the time remaining to them. Improved psychiatric and medical care for those who are terminally ill offer significant possibilities for suicide prevention.


Total iron-overload measurement in the human liver region by the magnetic iron detector.
An accurate assessment of body iron accumulation is essential for the diagnosis and therapy of iron overload in diseases, such as hemochromatosis, thalassemia and other forms of severe anemias. The magnetic iron detector (MID) is a room-temperature susceptometer, which measures the total iron overload in the liver. Since February 2005, about 600 patients have been assessed using this device. The iron overload is obtained by calculating the difference between the measured magnetization signal of the patient and the patient's background signal. The latter is the magnetization signal that the patient would generate with normal iron content. This study presents the method for calculating the background signal of healthy volunteers and the application of the same method to patients with iron burden in order to evaluate their overload. The present MID sensitivity is 0.8 g and the reproducibility of the iron overload measurement of the same patients is lower than 0.5 g. The MID does not require calibration with liver biopsies. We correlated the MID measurements with the results of 26 biopsies (R = 0.62), 64 superconducting quantum interference device susceptometer measurements (R = 0.79), 666 serum ferritin concentration measurements (R = 0.72), and 41 MRI- R2* measurements (R = 0.71).


Pseudomonas aeruginosa in cystic fibrosis: role of mucC in the regulation of alginate production and stress sensitivity.
Alginate production in Pseudomonas aeruginosa and the associated mucoid phenotype of isolates from cystic fibrosis patients are under the control of the algU mucABCD cluster. This group of genes encodes AlgU, the P. aeruginosa equivalent of the extreme heat shock sigma factor sigma E in Gram-negative bacteria, the AlgU-cognate anti-sigma factor MucA, the periplasmic protein MucB and a serine protease homologue, MucD. While mucA, mucB or mucD act as negative regulators of AlgU, the function of mucC is not known. In this study the role of mucC in P. aeruginosa physiology and alginate production has been addressed. Insertional inactivation of mucC in the wild-type P. aeruginosa strain PAO1 did not cause any overt effects on alginate synthesis. However, it affected growth of P. aeruginosa under conditions of combined elevated temperature and increased ionic strength or osmolarity. Inactivation of mucC in mucA or mucB mutant backgrounds resulted in a mucoid phenotype when the cells were grown under combined stress conditions of elevated temperature and osmolarity. Each of the stress factors tested separately did not cause comparable effects. The combined stress factors were not sufficient to cause phenotypically appreciable enhancement of alginate production in mucA or mucB mutants unless mucC was also inactivated. These findings support a negative regulatory role of mucC in alginate production by P. aeruginosa, indicate additive effects of muc genes in the regulation of mucoidy in this organism and suggest that multiple stress signals and recognition systems participate in the regulation of algU-dependent functions.


Mechanistic dichotomy in CpRu(CH(3)CN)(3)PF(6) catalyzed enyne cycloisomerizations.
Enynes are easily accessible building blocks as a result of the rich chemistry of alkynes and thus represent attractive substrates for ring formation. A ruthenium catalyst for cycloisomerization effects such reaction of 1,6- and 1,7-enynes typically at room temperature in acetone or DMF under neutral conditions. The reaction is effective for forming five- and six-membered rings of widely divergent structure. The alkyne may bear both election-donating and election-withdrawing substituents. The alkene may be di- or trisubstituted. Introduction of a quaternary center at the propargylic position of an ynoate, however, completely changes the nature of the reaction. In the case of a 1,6-enynoate, a seven-membered ring forms in excellent yield under equally mild conditions. Evidence is presented to indicate a complete change in mechanism. In the former case, the reaction involves the intermediacy of a ruthenacyclopentene. In the latter case, a C-H insertion to form a pi-allylruthenium intermediate is proposed and supported by deuterium-labeling studies. A rationale is presented for the structural dependence of the mechanism.


Reversion of an S49 cell cyclic AMP-dependent protein kinase structural gene mutant occurs primarily by functional elimination of mutant gene expression.
The regulatory subunits of cyclic AMP (cAMP)-dependent protein kinase from a dibutyryl cAMP-resistant S49 mouse lymphoma cell mutant, clone U200/65.1, and its revertants were visualized by two-dimensional polyacrylamide gel electrophoresis. Clone U200/65.1 co-expressed electrophoretically distinguishable mutant and wild-type subunits (Steinberg et al., Cell 10:381-391, 1977). In all 48 clones examined, reversion of the mutant to dibutyryl cAMP sensitivity was accompanied by alterations in regulatory subunit labeling patterns. Some spontaneous (3 of 11) and N-methyl-N'-nitro-N-nitrosoguanidine-induced (2 of 11) revertants retained mutant subunits, but these were altered in charge, degree of phosphorylation, or both. The charge alterations were consistent with single amino acid substitutions, suggesting that reversion was the result of second-site mutations in the mutant regulatory subunit allele that restored wild-type function, although not wild-type structure, to the gene product. The majority of spontaneous (8 of 11) and N-methyl-N'-nitro-N-nitrosoguanidine-induced (9 of 11) revertants and all of the revertants induced by ethyl methane sulfonate (14 of 14) and ICR191 (12 of 12) displayed only wild-type subunits. Dibutyryl cAMP-resistant mutants isolated from several of these revertants displayed new mutant but not wild-type subunits, suggesting that the revertant parent expresses only a single, functional regulatory subunit allele. The mutant regulatory subunit allele can, therefore, be modified in two general ways to produce revertant phenotypes: (i) by mutations that restore its wild-type function, and (ii) by mutations that eliminate its function.


Intralesional bleomycin injections for the palliation of epistaxis in hereditary hemorrhagic telangiectasia.
Epistaxis is a common complication in patients with hereditary hemorrhagic telangiectasia. Its treatment is generally aimed at controlling the frequency and severity of nasal hemorrhage and involves surgery, transcatheter embolization, topical treatment, or a combination. Despite this multitude of treatment methods, the long-term prognosis for many patients remains poor. We describe a patient in whom direct intralesional injections of bleomycin successfully palliated severe recurrent epistaxis for almost 2 years without the need for adjuvant therapy.


Differential activation of mitogen-activated protein kinases in AGS gastric epithelial cells by cag+ and cag- Helicobacter pylori.
The aim of this study was to determine whether Helicobacter pylori activates mitogen-activated protein (MAP) kinases in gastric epithelial cells. Infection of AGS cells with an H. pylori cag+ strain rapidly (5 min) induced a dose-dependent activation of extracellular signal-regulated kinases (ERK), p38, and c-Jun N-terminal kinase (JNK) MAP kinases, as determined by Western blot analysis and in vitro kinase assay. Compared with cag+ strains, cag- clinical isolates were less potent in inducing MAP kinase, particularly JNK and p38, activation. Isogenic inactivation of the picB region of the cag pathogenicity island resulted in a similar loss of JNK and p38 MAP kinase activation. The specific MAP kinase inhibitors, PD98059 (25 microM; MAP kinase kinase (MEK-1) inhibitor) and SB203580 (10 microM; p38 inhibitor), reduced H. pylori-induced IL-8 production in AGS cells by 78 and 82%, respectively (p < 0.01 for each). Both inhibitors together completely blocked IL-8 production (p < 0.001). However, the MAP kinase inhibitors did not prevent H. pylori-induced IkappaBalpha degradation or NF-kappaB activation. Thus, H. pylori rapidly activates ERK, p38, and JNK MAP kinases in gastric epithelial cells; cag+ isolates are more potent than cag- strains in inducing MAP kinase phosphorylation and gene products of the cag pathogenicity island are required for maximal MAP kinase activation. p38 and MEK-1 activity are required for H. pylori-induced IL-8 production, but do not appear to be essential for H. pylori-induced NF-kappaB activation. Since MAP kinases regulate cell proliferation, differentiation, programmed death, stress, and inflammatory responses, activation of gastric epithelial cell MAP kinases by H. pylori cag+ strains may be instrumental in inducing gastroduodenal inflammation, ulceration, and neoplasia.


The effectiveness of seasonal trivalent inactivated influenza vaccine in preventing laboratory confirmed influenza hospitalisations in Auckland, New Zealand in 2012.
Few studies report the effectiveness of trivalent inactivated influenza vaccine (TIV) in preventing hospitalisation for influenza-confirmed respiratory infections. Using a prospective surveillance platform, this study reports the first such estimate from a well-defined ethnically diverse population in New Zealand (NZ). A case test-negative design was used to estimate propensity adjusted vaccine effectiveness. Patients with a severe acute respiratory infection (SARI), defined as a patient of any age requiring hospitalisation with a history of a fever or a measured temperature ≥38°C and cough and onset within the past 7 days, admitted to public hospitals in South and Central Auckland were eligible for inclusion in the study. Cases were SARI patients who tested positive for influenza, while non-cases (controls) were SARI patients who tested negative. Results were adjusted for the propensity to be vaccinated and the timing of the influenza season. The propensity and season adjusted vaccine effectiveness (VE) was estimated as 39% (95% CI 16;56). The VE point estimate against influenza A (H1N1) was lower than for influenza B or influenza A (H3N2) but confidence intervals were wide and overlapping. Estimated VE was 59% (95% CI 26;77) in patients aged 45-64 years but only 8% (-78;53) in those aged 65 years and above. Prospective surveillance for SARI has been successfully established in NZ. This study for the first year, the 2012 influenza season, has shown low to moderate protection by TIV against influenza positive hospitalisation.


[Effect of intravenous hypnotics on autonomic response in patients with ischemic heart disease].
The purpose of the case study was to investigate the effect from the bolus dosing of midazolam and ketamine on the autonomic hemostasis in patients with ischemic heart disease (IHD) with the prevalence of the activity of sympathetic or parasympathetic parts in the autonomic nervous system. The IHD patients (n = 75) were examined, after scheduled preoperative therapy, active orthostatic tests and tests of cardiac rate variability (CRV to determinine a type of autonomic response. After orthostatic-test results were analyzed, patients with sympathicotonic (n = 15) and vagotonic (n = 15) types of vegetative response were selected. After midazolam at 0.08-0.12 mg/kg and ketamine at 0.5-1.3 mg/kg were administered, CRV was analyzed in the operating theater.


[Efficiency of feedback selection for recombination in Drosophila].
A number of theoretical models have been developed in population genetics to explain the evolution of recombination system under natural selection for adaptive traits. Using Drosophila cage populations of large size the effect of selection for resistance to daily temperature fluctuations on the rec-system was studied in this work. It is established that selection for population adaptability to extreme conditions leads during several tens of generations to significant increase in crossing-over rate within various parts of the genome (especially in b-cn and ru-h of chromosomes 2 and 3), as well as to relaxation of exchange interference for adjacent and non-adjacent segments. The Fisher's theoretical prognosis about tightening of linkage in optimal constant conditions was experimentally confirmed for the first time.


Patient safety in trauma: maximal impact management errors at a level I trauma center.
The Division of Research at JCAHO developed a taxonomy (common terminology and classification schema) to promote consistency in reporting and facilitate root cause analysis. We undertook a review of trauma management errors at our institution with maximal impact (death). The analysis was based on the Joint Commission on Accreditation of Healthcare Organizations (JCAHO) taxonomy. Trauma deaths between 2001 and 2006 at our Level I trauma were peer-reviewed to identify errors in management. The errors are classified according to type, domain, and cause. Seventy-six (9.9%) of 764 deaths had management errors contributing to potentially preventable deaths in 60 (errors in management might have contributed to death) and preventable deaths (management errors definitely contributed to death) in 16 patients. Questionable resuscitation was the commonest type and involved poor treatment in the majority. Errors were made in all domains but most commonly in the emergency department and the operating room and in the resuscitative phase. Human errors predominated. Management errors in the basics of trauma care continue even in established trauma centers, despite guidelines, protocols, and continuous performance improvement. Standardized reporting such as the taxonomy may result in progressive collection of patient safety data and lead to innovations to minimize these errors.


Anterior crucial ligament rupture: self-healing through dynamic intraligamentary stabilization technique.
Surgery involving arthroscopic reconstruction of the injured ligament is the gold standard treatment for torn anterior cruciate ligament (ACL). Recent studies support the hypothesis of biological self-healing of ruptured ACL. The aim of the study is to evaluate, in an animal model, the efficacy of a new technique, dynamic intraligamentary stabilization that utilizes biological self-healing for repair of acute ACL ruptures. The ACL in 11 adult female white alpine sheep was transected and in 8 sheep reconstructed by dynamic intraligamentary stabilization. To enhance the healing potential, microfracturing and collagen were used in all animals. The contralateral, non-operated knees served as controls. At 3 months postkilling, all animals were submitted to magnetic resonance imaging and biomechanical and histological evaluation. No surgery-related complications were observed. Postoperatively, all animals regularly used the operated leg with full weight bearing and no lameness. At the time of killing, all animals exhibited radiological and histological healing of the transacted ACL. Biomechanical tests confirmed successful restoration of anteroposterior translation in the dynamic intraligamentary stabilization knees. Histological examination revealed dense scar tissue at the ends of the transected ligaments exhibiting hypercellularity and hypervascularization. The dynamic intraligamentary stabilization technique successfully induced self-healing of ruptured ACL in a sheep model. Knee joints remained stable during the healing period allowing free range of motion and full weight bearing, and no signs of osteoarthritis or other intraarticular damage in the follow up were observed.


A universal TagModule collection for parallel genetic analysis of microorganisms.
Systems-level analyses of non-model microorganisms are limited by the existence of numerous uncharacterized genes and a corresponding over-reliance on automated computational annotations. One solution to this challenge is to disrupt gene function using DNA tag technology, which has been highly successful in parallelizing reverse genetics in Saccharomyces cerevisiae and has led to discoveries in gene function, genetic interactions and drug mechanism of action. To extend the yeast DNA tag methodology to a wide variety of microorganisms and applications, we have created a universal, sequence-verified TagModule collection. A hallmark of the 4280 TagModules is that they are cloned into a Gateway entry vector, thus facilitating rapid transfer to any compatible genetic system. Here, we describe the application of the TagModules to rapidly generate tagged mutants by transposon mutagenesis in the metal-reducing bacterium Shewanella oneidensis MR-1 and the pathogenic yeast Candida albicans. Our results demonstrate the optimal hybridization properties of the TagModule collection, the flexibility in applying the strategy to diverse microorganisms and the biological insights that can be gained from fitness profiling tagged mutant collections. The publicly available TagModule collection is a platform-independent resource for the functional genomics of a wide range of microbial systems in the post-genome era.


Safety and activity of alectinib against systemic disease and brain metastases in patients with crizotinib-resistant ALK-rearranged non-small-cell lung cancer (AF-002JG): results from the dose-finding portion of a phase 1/2 study.
Patients with non-small-cell lung cancer (NSCLC) and ALK rearrangements generally have a progression-free survival of 8-11 months while on treatment with the ALK inhibitor crizotinib. However, resistance inevitably develops, with the brain a common site of progression. More potent ALK inhibitors with consistently demonstrable CNS activity and good tolerability are needed urgently. Alectinib is a novel, highly selective, and potent ALK inhibitor that has shown clinical activity in patients with crizotinib-naive ALK-rearranged NSCLC. We did a phase 1/2 study of alectinib to establish the recommended phase 2 dose of the drug and examine its activity in patients resistant or intolerant to crizotinib. We enrolled patients with ALK-rearranged NSCLC who progressed on or were intolerant to crizotinib. We administered various oral doses of alectinib (300-900 mg twice a day) during the dose-escalation portion of the study (phase 1), to ascertain the recommended dose for phase 2. We used Response Evaluation Criteria in Solid Tumors criteria (version 1.1) to investigate the activity of alectinib in all patients with a baseline scan and at least one post-treatment scan (CT or MRI), with central radiological review of individuals with brain metastases. We assessed safety in all patients who received at least one dose of alectinib. Here, we present data for the phase 1 portion of the study, the primary objective of which was to establish the recommended phase 2 dose; phase 2 is ongoing. This trial is registered at ClinicalTrials.gov, number NCT01588028. 47 patients were enrolled. Alectinib was well tolerated, with the most common adverse events being fatigue (14 [30%]; all grade 1-2), myalgia (eight [17%]; all grade 1-2), and peripheral oedema (seven [15%] grade 1-2, one [2%] grade 3). Dose-limiting toxic effects were recorded in two patients in the cohort receiving alectinib 900 mg twice a day; one individual had grade 3 headache and the other had grade 3 neutropenia. The most common grade 3-4 adverse events were increased levels of γ-glutamyl transpeptidase (two [4%]), a reduction in the number of neutrophils (two [4%]), and hypophosphataemia (two [4%]). Three patients reported four grade 4 serious adverse events that were deemed unrelated to alectinib: acute renal failure; pleural effusion and pericardial effusion; and brain metastasis. At data cut-off (median follow-up 126 days [IQR 84-217]), 44 patients could be assessed for activity. Investigator-assessed objective responses were noted in 24 (55%) patients, with a confirmed complete response in one (2%), a confirmed partial response in 14 (32%), and an unconfirmed partial response in nine (20%). 16 (36%) patients had stable disease; the remaining four (9%) had progressive disease. Of 21 patients with CNS metastases at baseline, 11 (52%) had an objective response; six (29%) had a complete response (three unconfirmed) and five (24%) had a partial response (one unconfirmed); eight (38%) patients had stable disease and the remaining two (10%) had progressive disease. Pharmacokinetic data indicated that mean exposure (AUC0-10) after multiple doses of alectinib (300-600 mg twice a day) was dose-dependent. Alectinib was well tolerated, with promising antitumour activity in patients with ALK-rearranged NSCLC resistant to crizotinib, including those with CNS metastases. On the basis of activity, tolerability, and pharmacokinetic data, we chose alectinib 600 mg twice a day as the recommended dose for phase 2. Chugai Pharmaceuticals, F Hoffmann La-Roche.


Activation of the phospholipid scramblase TMEM16F by nanosecond pulsed electric fields (nsPEF) facilitates its diverse cytophysiological effects.
Nanosecond pulsed electric fields (nsPEF) are emerging as a novel modality for cell stimulation and tissue ablation. However, the downstream protein effectors responsible for nsPEF bioeffects remain to be established. Here we demonstrate that nsPEF activate TMEM16F (or Anoctamin 6), a protein functioning as a Ca2+-dependent phospholipid scramblase and Ca2+-activated chloride channel. Using confocal microscopy and patch clamp recordings, we investigated the relevance of TMEM16F activation for several bioeffects triggered by nsPEF, including phosphatidylserine (PS) externalization, nanopore-conducted currents, membrane blebbing, and cell death. In HEK 293 cells treated with a single 300-ns pulse of 25.5 kV/cm, Tmem16f expression knockdown and TMEM16F-specific inhibition decreased nsPEF-induced PS exposure by 49 and 42%, respectively. Moreover, the Tmem16f silencing significantly decreased Ca2+-dependent chloride channel currents activated in response to the nanoporation. Tmem16f expression also affected nsPEF-induced cell blebbing, with only 20% of the silenced cells developing blebs compared with 53% of the control cells. This inhibition of cellular blebbing correlated with a 25% decrease in cytosolic free Ca2+ transient at 30 s after nanoporation. Finally, in TMEM16F-overexpressing cells, a train of 120 pulses (300 ns, 20 Hz, 6 kV/cm) decreased cell survival to 34% compared with 51% in control cells (*, p < 0.01). Taken together, these results indicate that TMEM16F activation by nanoporation mediates and enhances the diverse cellular effects of nsPEF.


Hypolipidemic effects of nimesulide and celecoxib in experimentally induced hypercholesterolemia in rabbits.
Hypercholesterolemia plays an important role in the development of atherosclerotic disease, which is one of the leading causes of mortality in the world. Previous studies showed that cyclooxygenase (COX) inhibitors could be used in treating hypercholesterolemia. Thepresentstudywas designed to testwhether COX-2 inhibition can improve lipid profiles inhypercholesterolemia. Rabbits were fed a high-cholesterol diet to produce hypercholesterolemia. The role of COX-2 was evaluated using celecoxib and nimesulide. Rabbits were divided into 4 groups: the first with normal healthy rabbits, second with high-cholesterol diet and pretreatment with saline, third with high-cholesterol diet and pretreatment with celecoxib, and fourth with high-cholesterol diet and pretreatment with nimesulide. Total cholesterol, high-density lipoprotein (HDL), low-density lipoprotein (LDL), and triglyceride levels were measured at 2-week intervals in all the groups. Results show significantly high levels of serum cholesterol, LDL, and triglyceride but low levels of HDL in hypercholesterolemic rabbits pretreated with saline. Rabbits pretreated with nimesulide and celecoxib showed improvement compared to the saline-treated group. Improvement of lipid profile by celecoxib in hypercholesterolemic rabbits indicates the detrimental role of COX-2 during atherogenesis. However, the observed effects of nimesulide and celecoxib in hypercholesterolemia may be independent of their ability to inhibit COX-2. Nevertheless, both celecoxib and nimesulide show lipid-lowering potential in experimental hypercholesterolemia.


Point-of-Care Coagulation Testing in Cardiac Surgery.
Bleeding complications after cardiac surgery are common and are associated with increased morbidity and mortality. Their etiology is multifactorial, and treatment decisions are time sensitive. Point-of-care (POC) testing has an advantage over standard laboratory tests for faster turn-around times, and timely decision on coagulation intervention(s). The most common POC coagulation testing is the activated clotting time (ACT), used to monitor heparin therapy while on cardiopulmonary bypass. Viscoelastic coagulation tests including thromboelastometry (ROTEM) and thromboelastography (TEG) have been recommended for the treatment of postoperative bleeding after cardiac surgery because the ROTEM/TEG-guided treatment algorithms reduced the use of blood products. Other POC tests are commercially available, but there is sparse evidence for their routine use in cardiac surgery. These devices include heparin management systems, POC prothrombin time and activated partial thromboplastin time, POC fibrinogen assay, and whole blood platelet function tests. There are multiple confounding elements and conditions associated with cardiac surgery, which can significantly alter test results. Anemia and thrombocytopenia are regularly associated with deviations in many POC devices. In summary, POC coagulation testing allows for rapid clinical decisions in hematological interventions, and, when used in conjunction with a proper transfusion algorithm, may reduce blood product usage, and potentially complications associated with blood transfusion.


Increased expression of MMP-2, MMP-9 (type IV collagenases/gelatinases), and MT1-MMP in canine X-linked Alport syndrome (XLAS).
Alport syndrome is a group of genetic disorders resulting from mutations in either the alpha3(IV), alpha4(IV) or alpha5(IV) collagen chains. The disease is characterized by a progressive glomerulonephritis, usually associated with a high-frequency specific sensorineural hearing loss, dot and fleck retinopathy, and lens abnormalities. Dogs with naturally occurring genetic disorders of basement membrane collagen (type IV) may serve as animal models of Alport syndrome. In this study, a well-characterized naturally occurring canine model was employed to demonstrate a potential role for matrix metalloproteinases (MMPs) in Alport renal disease pathogenesis. Adolescent male dogs that developed renal failure were euthanized and necropsied. Clinicopathologic features of the disease were characterized, and kidneys from normal and Alport dogs were analyzed by gelatin zymography, Western blotting, in situ zymography, immunohistology, and by reverse transcription polymerase chain reaction (RT-PCR) for expression of MMP-2, MMP-9, and membrane type 1-MMP (MT1-MMP). Affected dogs developed proteinuria and rapidly progressive juvenile-onset chronic renal failure. The activities of MMP-2 and MMP-9 were significantly induced in Alport kidney. In situ zymography confirmed elevated active metalloproteinases in kidney cryosections of affected dogs. The mRNAs encoding MMP-2, MMP-9 and MT1-MMP were also increased in Alport dogs suggesting that elevated expression of MMPs reflects events in the progression of Alport syndrome in dogs. Elevated expression of MMP-2, MMP-9, and MT1-MMP is observed in fibrotic renal cortex from X-linked Alport syndrome dogs. These findings suggest that MMPs may play an important role in matrix accumulation associated with progressive renal scarring in this model.


Prostate Cancer--CHI Conference. Advances in understanding diagnostics and therapy.
As the title indicates, the purpose of this meeting was to highlight new information relative to the etiology, diagnosis and potential therapy of prostate cancer. Prostate cancer is the most prevalent internal cancer affecting men in the USA and the second most frequent cause of cancer-related deaths. Ongoing studies are identifying potentially important genes and target molecules involved in prostate cancer. These reagents will permit improved diagnostics and offer the promise of enhanced therapeutic targets for this cancer. The meeting was divided into four general areas. Approximately 70 scientists representing academic, governmental and industrial sectors attended this meeting.


Nipah virus sequesters inactive STAT1 in the nucleus via a P gene-encoded mechanism.
The Nipah virus (NiV) phosphoprotein (P) gene encodes the C, P, V, and W proteins. P, V, and W, have in common an amino-terminal domain sufficient to bind STAT1, inhibiting its interferon (IFN)-induced tyrosine phosphorylation. P is also essential for RNA-dependent RNA polymerase function. C is encoded by an alternate open reading frame (ORF) within the common amino-terminal domain. Mutations within residues 81 to 113 of P impaired its polymerase cofactor function, as assessed by a minireplicon assay, but these mutants retained STAT1 inhibitory function. Mutations within the residue 114 to 140 region were identified that abrogated interaction with and inhibition of STAT1 by P, V, and W without disrupting P polymerase cofactor function. Recombinant NiVs were then generated. A G121E mutation, which abrogated inhibition of STAT1, was introduced into a C protein knockout background (C(ko)) because the mutation would otherwise also alter the overlapping C ORF. In cell culture, relative to the wild-type virus, the C(ko) mutation proved attenuating but the G121E mutant virus replicated identically to the C(ko) virus. In cells infected with the wild-type and C(ko) viruses, STAT1 was nuclear despite the absence of tyrosine phosphorylation. This latter observation mirrors what has been seen in cells expressing NiV W. In the G121E mutant virus-infected cells, STAT1 was not phosphorylated and was cytoplasmic in the absence of IFN stimulation but became tyrosine phosphorylated and nuclear following IFN addition. These data demonstrate that the gene for NiV P encodes functions that sequester inactive STAT1 in the nucleus, preventing its activation and suggest that the W protein is the dominant inhibitor of STAT1 in NiV-infected cells.


The effects of transecting the IXth and Xth cranial nerves on hydromineral balance in the eel Anguilla anguilla.
The IXth and the Xth cranial nerves in Anguilla anguilla were transected, and the effects upon ion and water balance were studied in fresh water and sea water, and during transfer from fresh water and vice versa. In fresh water there is a slow demineralization due to an excess loss of Na and Cl ions. During freshwater to seawater transfer the eel survives only for 4-5 days. The fish do not drink and Na efflux does not increase enough to extrude excess ions. In sea water the glossopharyngeal and vagus nerves are necessary for the maintenance of the hydromineral balance. Denervation is followed by an increase in plasma ion concentrations. Na fluxes are not modified and increased water loss is not compensated by drinking. The rapid reduction of Na efflux during transfer from sea water to fresh water is not modified by denervation.


Identification of a novel testis-specific leucine-rich protein in humans and mice.
A novel testis-specific protein, termed LRTP, was identified by screening both human and mouse testis and mouse pachytene spermatocyte cDNA libraries. Sequence analyses (GenBank accession number: AF092208) revealed that LRTP contains an amino terminus leucine-rich repeat domain. There are several acidic regions rich in glutamic acid in the C-terminus. The sequence, by GenBank search, shows similarities to LANP and SDS22+, leucine-rich repeat proteins localized to the nucleus and involved in the regulation of protein phosphatases. In mouse, the mRNA is first detected at about Day 14 postpartum, presumably when mid-pachytene spermatocytes are first seen. In situ hybridization confirmed the expression of the LRTP mRNA at this stage of spermatogenesis. Immunohistochemical analysis revealed that the protein is most abundant in the cytoplasm of pachytene and diplotene cells, corresponding to late prophase of meiosis I. Immunohistochemical localization is markedly reduced in secondary spermatocytes, suggesting a functional association of LRTP with meiosis. An LRTP cDNA probe did not bind to mouse ovary RNA in a dot blot assay.


Multi-photon near-infrared emission saturation nanoscopy using upconversion nanoparticles.
Multiphoton fluorescence microscopy (MPM), using near infrared excitation light, provides increased penetration depth, decreased detection background, and reduced phototoxicity. Using stimulated emission depletion (STED) approach, MPM can bypass the diffraction limitation, but it requires both spatial alignment and temporal synchronization of high power (femtosecond) lasers, which is limited by the inefficiency of the probes. Here, we report that upconversion nanoparticles (UCNPs) can unlock a new mode of near-infrared emission saturation (NIRES) nanoscopy for deep tissue super-resolution imaging with excitation intensity several orders of magnitude lower than that required by conventional MPM dyes. Using a doughnut beam excitation from a 980 nm diode laser and detecting at 800 nm, we achieve a resolution of sub 50 nm, 1/20th of the excitation wavelength, in imaging of single UCNP through 93 μm thick liver tissue. This method offers a simple solution for deep tissue super resolution imaging and single molecule tracking.


Label-free quantification of host cell protein impurity in recombinant hemoglobin materials.
Quantitative analysis relies on pure-substance primary calibrators with known mass fractions of impurity. Here, label-free quantification (LFQ) is being evaluated as a readily available, reliable method for determining the mass fraction of host cell proteins (HCPs) in bioengineered proteins which are intended for use as protein calibration standards. In this study a purified hemoglobin-A2 (HbA2) protein, obtained through its overexpression in E. coli, was used. Two different materials were produced: natural and U15N-labeled HbA2. For the quantification of impurities, precursor ion (MS1-) intensities were integrated over all E. coli proteins identified and divided by the intensities obtained for HbA2. This ratio was calibrated against the corresponding results for an E. coli cell lysate, which had been spiked at known mass ratios to pure HbA2. To demonstrate the universal applicability of LFQ, further proteomes (yeast and human K562) were then alternatively used for calibration and found to produce comparable results. Valid results were also obtained when the complexity of the calibrator was reduced to a mix of just nine proteins, and a minimum of five proteins was estimated to be sufficient to keep the sampling error below 15%. For the studied materials, HbA2 mass fractions (or purities) of 923 and 928 mg(HbA2)/g(total protein) were found with expanded uncertainties (U) of 2.8 and 1.3%, resp. Value assignment by LFQ thus contributes up to about 3% of the overall uncertainty of HbA2 quantification when these materials are used as calibrators. Further purification of the natural HbA2 yielded a mass fraction of 999.1 mg/g, with a negligible uncertainty (U = 0.02%), though at a significant loss of material. If an overall uncertainty of 5% is acceptable for protein quantification, working with the original materials would therefore definitely be viable, circumventing the need of further purification.


Detection of clostridium difficile antigen and toxin in stool specimens: Comparison of the C. difficile quik chek complete enzyme immunoassay and GeneXpert C. difficile polymerase chain reaction assay.
Accurate and rapid laboratory diagnosis of Clostridium difficile infections (CDI) remains a significant challenge. A two-step algorithm for detection of toxigenic C. difficile in stool based on initial screening for glutamate dehydrogenase assay followed by confirmation by toxin A+B detection using an enzyme immunoassay (EIA) or molecular assay has been proposed. We aimed to evaluate the C. difficile Quik Chek Complete® (QCC-EIA) versus the GeneXpert® C. difficile polymerase chain reaction (PCR) assay in this two-step algorithm. Two hundred and ten liquid stool samples obtained between June 2014 and June 2015 from patients suspected of CDI were tested by the QCC-EIA and GeneXpert PCR assay. The GeneXpert assay was used as the reference standard to calculate the QCC-EIA sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV). Of the 210 stool samples tested, 43 (20.5%) were positive by QCC-EIA, while 31 (14.8%) were positive by GeneXpert assay. The sensitivity and specificity of the QCC-EIA were found to be 100 and 93%, respectively; the PPV and NPV were 72 and 100%, respectively. The binary toxin was detected in 12 (38.7%) and tcdC gene deletion in 3 (9.6%). The low specificity of QCC-EIA makes it less reliable as a confirmatory test for CDI diagnosis. This test may be used as a screening test in a two-step algorithm when combined with a molecular assay or another confirmatory test.


Novel pandemic influenza A (H1N1) virus infection modulates apoptotic pathways that impact its replication in A549 cells.
It is not well-known whether apoptosis signaling affects influenza virus infection and reproduction in human lung epithelial cells. Using A549 cell line, we studied the relationship of some apoptosis-associated molecules with novel pandemic influenza A (H1N1) virus, A/California/04/2009. Infected cells displayed upregulated Fas ligand, activated FADD and caspase-8, and downregulated FLIP in the extrinsic apoptotic pathway. p53 expression increased and Bcl-XL expression decreased in the intrinsic pathway. Expression of pre-apoptotic molecules (FasL, FADD, and p53) increased virus replication, while inhibition of activity of FADD, caspase-8 and caspase-3, and expression of anti-apoptotic proteins (FLIP and Bcl-XL) decreased virus replication. p38, ERK and JNK from MAPK pathways were activated in infected cells, and inhibition with their inhibitors diminished virus replication. In the p38 superfamily, p38α expression increased viral RNA production, while expression of p38β and p38γ decreased. These data indicated that influenza virus induces apoptotic signaling pathways, which benefit virus replication.


Role of PDGF-A expression in the control of vascular smooth muscle cell growth by transforming growth factor-beta.
Transforming growth factor-beta (TGF-beta) is a multifunctional regulatory peptide that can inhibit or promote the proliferation of cultured vascular smooth muscle cells (SMCs), depending on cell density (Majack, R. A. 1987. J. Cell Biol. 105:465-471). In this study, we have examined the mechanisms underlying the growth-promoting effects of TGF-beta in confluent SMC cultures. In mitogenesis assays using confluent cells, TGF-beta was found to potentiate the stimulatory effects of serum, PDGF, and basic fibroblast growth factor (bFGF), and was shown to act individually as a mitogen for SMC. In gene and protein expression experiments, TGF-beta was found to regulate the expression of PDGF-A and thrombospondin, two potential mediators of SMC proliferative events. The induction of thrombospondin protein and mRNA was density-dependent, delayed relative to its induction by PDGF and, based on cycloheximide experiments, appeared to depend on the de novo synthesis of an intermediary protein (probably PDGF-A). The relationship between PDGF-A expression and TGF-beta-mediated mitogenesis was investigated, and it was determined that a PDGF-like activity (probably PDGF-A) was the biological mediator of the growth-stimulatory effects of TGF-beta on confluent SMC. The effects of purified homodimers of PDGF-A on SMC replication were investigated, and it was determined that PDGF-AA was mitogenic for cultured SMC, particularly when used in combination with other growth factors such as bFGF and PDGF-BB. The data suggest several molecular mechanisms that may account for the ability of TGF-beta to promote the growth of confluent SMC in culture.


THERAPY OF ENDOCRINE DISEASE: Central neck dissection: a step forward in the treatment of papillary thyroid cancer.
Since its introduction in the '70s and '80s, CND for papillary cancer is here to stay. Compartment VI should always be explored during surgery for papillary thyroid carcinoma (PTC) for obvious lymph node metastases. These can be easily spotted by an experienced surgeon or, eventually, by frozen section. No doubt, obvious nodal disease in the Delphian, paratracheal and subithsmic areas should be dissected in a comprehensive manner (therapeutic central neck dissection), avoiding the selective removal of suspicious nodes. Available evidence for routine prophylactic CND is not completely satisfactory. Our group's opinion, however, is that it reduces or even eliminates the need for repeat surgery in the central neck, better defines the extent (and stage) of the disease and provides a further argument against routine radioiodine ablation. Thus, PTC is becoming more and more a surgical disease that can be cured by optimized surgery alone in the majority of cases. Prophylactic CND, however, involves a higher risk for the parathyroid function and should be skilfully performed, preferably only on the same side as the primary tumour and preserving the cervical portion of the thymus.


Assessing heterogeneity of treatment effect analyses in health-related cluster randomized trials: A systematic review.
Cluster-randomized trials (CRTs) are being increasingly used to test a range of interventions, including medical interventions commonly used in clinical practice. Policies created by the NIH and the Food and Drug Administration (FDA) require the reporting of demographics and the examination of demographic heterogeneity of treatment effect (HTE) for individually randomized trials. Little is known about how frequent demographics are reported and HTE analyses are conducted in CRTs. We sought to understand the prevalence of HTE analyses and the statistical methods used to conduct them in CRTs focused on treating cardiovascular disease, cancer, and chronic lower respiratory diseases. Additionally, we also report on the proportion of CRTs that reported on baseline demographics of its populations and conducted demographic HTE analyses. We searched PubMed and Embase for CRTs published between 1/1/2010 and 3/29/2016 that focused on treating the top 3 Center for Disease Control causes of death (cardiovascular disease, chronic lower respiratory disease, and cancer). Evidence Screening And Review: Of 1,682 unique titles, 117 abstracts were screened. After excluding 53 articles, we included 64 CRT publications and abstracted information on study characteristics and demographic information, statistical analysis, HTE analysis, and study quality. Age and sex were reported in greater than 95.3% of CRTs, while race and ethnicity were reported in only 20.3% of CRTs. HTE analyses were conducted in 28.1% (n = 18) of included CRTs and 77.8% (n = 12) were prespecified analyses. Four CRTs conducted a demographic subgroup analysis. Only 6/18 CRTs used interaction testing to determine whether HTE existed. Baseline demographic reporting was high for age and sex in CRTs, but was uncommon for race and ethnicity. HTE analyses were uncommon and was rare for demographic subgroups, which limits the ability to examine the extent of benefits or risks for treatments tested with CRT designs.